Biosynthesis of an Opine Metallophore by <i>Pseudomonas aeruginosa</i>

McFarlane, Jeffrey S.; Lamb, Audrey L.

doi:10.1021/acs.biochem.7b00804

Cited by 43 publications

(68 citation statements)

References 24 publications

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“…Staphylopine-like operons are found in other pathogenic species, including P. aeruginosa , in which the products of those genes are needed for growth within the host environment (188). Pseudopaline, the opine metallophore produced by these staphylopine-like operons, has recently been identified, and evidence that it plays a role in zinc and nickel import is available (189–191). …”

Section: Copper-binding Siderophores and Metallophoresmentioning

confidence: 99%

Chalkophores

Kenney

Rosenzweig

2018

Annu. Rev. Biochem.

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Copper-binding metallophores, or chalkophores, play a role in microbial copper homeostasis that is analogous to that of siderophores in iron homeostasis. The best-studied chalkophores are members of the methanobactin (Mbn) family—ribosomally produced, posttranslationally modified natural products first identified as copper chelators responsible for copper uptake in methane-oxidizing bacteria. To date, Mbns have been characterized exclusively in those species, but there is genomic evidence for their production in a much wider range of bacteria. This review addresses the current state of knowledge regarding the function, biosynthesis, transport, and regulation of Mbns. While the roles of several proteins in these processes are supported by substantial genetic and biochemical evidence, key aspects of Mbn manufacture, handling, and regulation remain unclear. In addition, other natural products that have been proposed to mediate copper uptake as well as metallophores that have biologically relevant roles involving copper binding, but not copper uptake, are discussed.

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Section: Copper-binding Siderophores and Metallophoresmentioning

confidence: 99%

Chalkophores

Kenney

Rosenzweig

2018

Annu. Rev. Biochem.

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“…In support of this idea, our data demonstrate that iron limitation results in increased levels of proteins encoded by the cnt operon, which mediate the synthesis, secretion, and uptake of a novel opine metallophore involved in zinc uptake ( Fig 7A-B ). Interestingly, CntL, the nicotianamine synthase (51), was not statistically increased by iron depletion, and CntI, the predicted cytoplasmic membrane exporter of pseudopaline (52), was not detected. Our finding that CntL protein levels are not affected by iron depletion is consistent with a recent study detailing the role of pseudopaline in zinc uptake (52).…”

Section: Resultsmentioning

confidence: 98%

Proteomic analysis of thePseudomonas aeruginosairon starvation response reveals PrrF sRNA-dependent regulation of amino acid metabolism, iron-sulfur cluster biogenesis, motility, and zinc homeostasis

Nelson

Huang

et al. 2018

Preprint

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Iron is a critical nutrient for most microbial pathogens, and the innate immune system exploits this requirement by sequestering iron and other metals through a process termed nutritional immunity. The opportunistic pathogen Pseudomonas aeruginosa provides a model system for understanding the microbial response to host iron depletion, as this organism exhibits a high requirement for iron as well as an exquisite ability to overcome iron deprivation during infection. Hallmarks of P. aeruginosa’s iron starvation response include the induction of multiple high affinity iron acquisition systems and an “iron sparing response” that is post-transcriptionally mediated by the PrrF small regulatory RNAs (sRNAs). Here, we used liquid chromatography-tandem mass spectrometry to conduct label-free proteomics of the P. aeruginosa iron starvation response, revealing several iron-regulated processes that have not been previously described. Iron starvation induced multiple proteins involved in branched chain and aromatic amino acid catabolism, providing the capacity for iron-independent entry of carbons into the TCA cycle. Proteins involved in sulfur assimilation and cysteine biosynthesis were reduced upon iron starvation, while proteins involved in iron-sulfur cluster biogenesis were paradoxically increased, highlighting the central role of iron in P. aeruginosa metabolism. Iron starvation also resulted in changes in the expression of several zinc-responsive proteins, as well as the first experimental evidence for increased levels of twitching motility proteins upon iron starvation. Subsequent proteomics analyses demonstrated that the PrrF sRNAs were required for iron regulation of many of these newly-identified proteins, and we identified PrrF complementarity with mRNAs encoding key iron-regulated proteins involved in amino acid metabolism, iron-sulfur biogenesis, and zinc homeostasis. Combined, these results provide the most comprehensive view of the P. aeruginosa iron starvation response to date and outline novel roles for the PrrF sRNAs in the P. aeruginosa iron sparing response and pathogenesis.AUTHOR SUMMARYIron is central for the metabolism of almost all microbial pathogens, and as such this element is sequestered by the host innate immune system to restrict microbial growth. Defining the response of microbial pathogens to iron starvation is therefore critical for understanding how pathogens colonize and propagate within the host. The opportunistic pathogen Pseudomonas aeruginosa, which causes significant morbidity and mortality in compromised individuals, provides an excellent model for studying this response due to its high requirement for iron yet well-documented ability to overcome iron starvation. Here we used label-free proteomics to investigate the P. aeruginosa iron starvation response, revealing a broad landscape of metabolic and metal homeostasis changes that have not previously been described. We further provide evidence that many of these processes are regulated through the iron responsive PrrF small regulatory RNAs, which are integral to P. aeruginosa iron homeostasis and virulence. These results demonstrate the power of proteomics for defining stress responses of microbial pathogens, and they provide the most comprehensive analysis to date of the P. aeruginosa iron starvation response.

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“…indicating that different StP-like chemicals are present in different microbes (18,21). Recent investigations in Pseudomonas aeruginosa uncovered a new nicotianamine-like metallophore pseudopaline that also mediates bacterial zinc acquisition under zinc-poor conditions (22)(23)(24). In contrast to the biosynthetic pathway of StP, the pseudopaline biosynthetic pathway lacks the histidine racemase and incorporates L-histidine and α-ketoglutarate instead of D-histidine and pyruvate into the scaffold of the metallophore (22)(23)(24).…”

Section: Significancementioning

confidence: 99%

Mechanistic insights into staphylopine-mediated metal acquisition

Song

Zhang

Chen

et al. 2018

Proc. Natl. Acad. Sci. U.S.A.

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Metal acquisition is vital to pathogens for successful infection within hosts. Staphylopine (StP), a broad-spectrum metallophore biosynthesized by the major human pathogen, , plays a central role in transition-metal acquisition and bacterial virulence. The StP-like biosynthesis loci are present in various pathogens, and the proteins responsible for StP/metal transportation have been determined. However, the molecular mechanisms of how StP/metal complexes are recognized and transported remain unknown. We report multiple structures of the extracytoplasmic solute-binding protein CntA from the StP/metal transportation system in apo form and in complex with StP and three different metals. We elucidated a sophisticated metal-bound StP recognition mechanism and determined that StP/metal binding triggers a notable interdomain conformational change in CntA. Furthermore, CRISPR/Cas9-mediated single-base substitution mutations and biochemical analysis highlight the importance of StP/metal recognition for StP/metal acquisition. These discoveries provide critical insights into the study of novel metal-acquisition mechanisms in microbes.

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Biosynthesis of an Opine Metallophore by Pseudomonas aeruginosa

Cited by 43 publications

References 24 publications

Chalkophores

Chalkophores

Proteomic analysis of thePseudomonas aeruginosairon starvation response reveals PrrF sRNA-dependent regulation of amino acid metabolism, iron-sulfur cluster biogenesis, motility, and zinc homeostasis

Mechanistic insights into staphylopine-mediated metal acquisition

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